Molecular Plant Biology and Biotechnology Lab, CSIR-Central Institute of Medicinal and Aromatic Plants, Research Centre, Bengaluru, 560065, India.
Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, 226015, India.
Plant J. 2022 Sep;111(5):1308-1323. doi: 10.1111/tpj.15890. Epub 2022 Jul 12.
Terpene synthases (TPSs) have diverse biological functions in plants. Though the roles of TPSs in herbivore defense are well established in many plant species, their role in bacterial defense has been scarce and is emerging. Through functional genomics, here we report the in planta role of potato (Solanum tuberosum) terpene synthase (StTPS18) in bacterial defense. Expression of StTPS18 was highest in leaves and was induced in response to Pseudomonas syringae and methyl jasmonate treatments. The recombinant StTPS18 exhibited bona fide (E,E)-farnesol synthase activity forming a sesquiterpenoid, (E,E)-farnesol as the sole product, utilising (E,E)-farnesyl diphosphate (FPP). Subcellular localization of GFP fusion protein revealed that StTPS18 is localized to the cytosol. Silencing and overexpression of StTPS18 in potato resulted in reduced and enhanced tolerance, respectively, to bacterial pathogens P. syringae and Ralstonia solanacearum. Bacterial growth assay using medium containing (E,E)-farnesol significantly inhibited P. syringae growth. Moreover, StTPS18 overexpressing transgenic potato and Nicotiana tabacum leaves, and (E,E)-farnesol and P. syringae infiltrated potato leaves exhibited elevated expression of sterol pathway and members of pathogenesis-related genes with enhanced phytosterol accumulation. Interestingly, enhanced phytosterols in C -(E,E)-farnesol infiltrated potato leaves were devoid of any noticeable C labeling, indicating no direct utilization of (E,E)-farnesol in phytosterols formation. Furthermore, leaves of StTPS18 overexpressing transgenic lines had no detectable (E,E)-farnesol similar to the control plant, and emitted lower levels of sesquiterpenes than the control. These findings point towards an indirect involvement of StTPS18 and its product (E,E)-farnesol in bacterial defense through upregulation of phytosterol biosynthesis and defense genes.
萜烯合酶(TPSs)在植物中具有多种生物学功能。尽管在许多植物物种中,TPSs 在食草动物防御中的作用已得到充分证实,但它们在细菌防御中的作用却很少见,并且正在出现。通过功能基因组学,我们在这里报告了马铃薯(Solanum tuberosum)萜烯合酶(StTPS18)在细菌防御中的体内作用。StTPS18 的表达在叶片中最高,并响应丁香假单胞菌和茉莉酸甲酯处理而被诱导。重组 StTPS18 表现出真正的(E,E)-法呢醇合酶活性,形成倍半萜烯(E,E)-法呢醇作为唯一产物,利用(E,E)-法呢基二磷酸(FPP)。GFP 融合蛋白的亚细胞定位表明 StTPS18 定位于细胞质。马铃薯中 StTPS18 的沉默和过表达分别导致对细菌病原体丁香假单胞菌和茄青枯菌的耐受性降低和增强。使用含有(E,E)-法呢醇的培养基进行细菌生长测定显著抑制了丁香假单胞菌的生长。此外,StTPS18 过表达转基因马铃薯和烟草叶片以及(E,E)-法呢醇和丁香假单胞菌渗透的马铃薯叶片表现出固醇途径和致病性相关基因成员的表达升高,并增强了植物甾醇积累。有趣的是,在 C-(E,E)-法呢醇渗透的马铃薯叶片中,增强的植物甾醇没有任何明显的 C 标记,表明(E,E)-法呢醇在植物甾醇形成中没有直接利用。此外,StTPS18 过表达转基因株系的叶片与对照植物一样,没有检测到(E,E)-法呢醇,并且比对照植物释放出较低水平的倍半萜烯。这些发现表明,StTPS18 及其产物(E,E)-法呢醇通过上调植物甾醇生物合成和防御基因,间接参与细菌防御。